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RESEARCH ON DENGUE IN TISSUE CULTURE : V. Production of Anti-Dengue Antibodies in Tissue Cultures from Monkeys Inoculated with an Attenuated Type I Dengue Virus Strain

English (英語)

The Kobe journal of the medical sciences

18(3)

143-152

1972-09

As a continuation of the previous studies (reported in this JOURNAL, 1966, 12, 199-205; Ibid, 1969, 15, 163-180), in vitro production of antibodies against dengue virus was investigated by means of tissue culture techniques. From monkeys inoculated with live dengue virus of human-attenuated strain (type 1, Mochizuki), tissues (spleen, lymph node, lung, liver, kidney and testis) were excised and cultivated by the plasma-embedded method. Culture fluids harvested at various times during cultivation were measured for neutralizing (NT) and hemagglutination-inhibiting (HI) titers against the original dengue virus. Sucrose density gradient centrifugation (Suc-DGC) and mercaptoethanol (ME) treatment were also performed to analyze the immunoglobulins. The anti-dengue NT and HI activities were clearly demonstrated in culture fluids of spleen, lymph node, lung or liver. The same activities of testis culture fluids were very low, and those of kidney cultures varied between different specimens. Under the same conditions, tissue culture fluids from monkeys inoculated with the virus killed by formalin showed few NT or HI activities. Of the HI-positive samples, the Suc-DGC profiles could be divided into two peaks, i.e., bottom fractions, corresponding to 19S globulins and ME-sensitive, and top fractions, corresponding to 7S globulins and ME-resistant. General tendencies were that the culture fluids harvested early after the beginning of cultivation possessed the two peaks, while those taken during later stages of cultivation had a single peak in the top fractions. The patterns were similar in the samples both from monkeys receiving a primary injection of virus and those receiving the booster injection. A tendency was noted, however, that the titers of the latter groups were higher than those of the former. Virologic and immunologic significances of the data obtained are discussed, referring to previous works relating to in vitro production of antibodies.

monkey

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